Titolo | Improved protocol for Agrobacterium mediated transformation of tomato and production of transgenic plants containing carotenoid biosynthetic gene CsZCD |
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Tipo di pubblicazione | Articolo su Rivista peer-reviewed |
Anno di Pubblicazione | 2007 |
Autori | Qiu, D., Diretto Gianfranco, Tavarza R., and Giuliano Giovanni |
Rivista | Scientia Horticulturae |
Volume | 112 |
Paginazione | 172-175 |
ISSN | 03044238 |
Parole chiave | agricultural research, Agrobacterium, Agrobacterium tumefaciens, bacterium, Carotenoid, Crocus, crop production, enzyme, fruit, gene, Lycopersicon esculentum, Rhizobium, transgenic plant |
Abstract | Improved protocol for Agrobacterium mediated transformation of tomato (Lycopersicon esculentum) Micro-Tom was developed to use in corporation of the carotenoid biosynthetic genes CsZCD (Crocus zeaxanthin 7,8-cleavage dioxygenase). From these experiment, a transformation methodology using explants from cotyledons cultured for 1 day on the medium with zeatin 2 mg/L, IAA 0.1 mg/L, carefully submerged in the Agrobacterium inoculum for 20 min, then concultured with the agrobacterium for 3 days on the same medium, followed by a transfer to the same medium with 500 mg/L cefotaxin for 3 days and then by a transfer to the same medium with 100 mg/L kanamycin and 500 mg/L carabenillin for 6-8 weeks and resulted in a greater than 20% transformation efficiency in the concentration of Agrobacterium OD600 = 0.2 tested. In this transformation method, no feeder layers were used and the subculture media was minimal. Among the Agrobacterium concentrations of OD600 = 0.2, 0.5 and 1.0, the best transformation efficiency, 20.87%, was obtained by using OD600 = 0.2, which was significantly higher than that of OD600 = 1.0. The presence of the inserted target genes was checked using a rapid and efficient PCR test. The protocol was successfully employed in the production of transgenic Micro-Tom tomato containing the carotenoid biosynthesis CsZCD under constructive promoter. This procedure represents a simple, efficient and general means of transforming tomato. © 2006 Elsevier B.V. All rights reserved. |
Note | cited By 32 |
URL | https://www.scopus.com/inward/record.uri?eid=2-s2.0-33847342467&doi=10.1016%2fj.scienta.2006.12.015&partnerID=40&md5=b677e54df27074820c40a6ef843b49db |
DOI | 10.1016/j.scienta.2006.12.015 |
Citation Key | Qiu2007172 |